Gramicidin S and Melittin - Potential anti-viral therapeutic peptides to treat SARS-CoV-2 infection

  1. Mohammed Ghalib
  2. Yash Parekh
  3. Sarena Banu
  4. Sushma Ram
  5. Ramakrishnan Nagaraj
  6. Bokara Kiran Kumar
  7. Mohammed M Idris

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Abstract

The COVID19 pandemic has resulted in multipronged approaches for treatment of the disease. Since de novo discovery of drugs is time consuming, repurposing of molecules is now considered as one of the alternative strategies to treat COVID19. Antibacterial peptides are being recognized as attractive candidates for repurposing to treat viral infections. In this study, we describe the anti-SARS-CoV-2 activity of gramicidin S and melittin peptides obtained from Bacillus brevis and bee venom respectively. Our in vitro antiviral assay results showed significant decrease in the viral load compared to the untreated group with no/very less cytotoxicity. The EC 50 values for gramicidin S and melittin are calculated as 1.571μg and 0.656μg respectively. Both the peptides treated to the SARS-CoV-2 infected Vero cells showed viral clearance from 12 hours onwards with a maximal clearance after 24 hours post infection. Based on proteome analysis it was found that more than 250 proteins were found to be differentially regulated in the gramicidin S and melittin treated SARS-CoV-2 infected Vero cells against control SARS-CoV-2 infected Vero cells after 24 and 48 hours post infection. The identified proteins were found to be associated in the metabolic and mRNA processing of the Vero cells post-treatment and infection. Both these peptides could be attractive candidates for repurposing to treat SARS-CoV-2 infection.

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  1. ScreenIT

    SciScore for 10.1101/2021.10.21.465254: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Later the cells were incubated with the secondary anti-Rabbit IgG antibody conjugated with Alexa Fluor 488 (Life Technologies, Cat. no.: A11008; Lot no.: 1735088) at the dilution of 1:200 in 1% BSA made in PBS. Rhodamine Phalloidin (Life Technologies, Cat. no.: R415; Lot no.: 1738179) was used to label F-actin.
    anti-Rabbit IgG
    suggested: (Thermo Fisher Scientific Cat# A-11008, RRID:AB_143165)
    F-actin
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    After the incubation, virus inoculum containing melittin & gramicidin S was added to the Vero cells in duplicates (50μl /well).
    Vero
    suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
    Software and Algorithms
    SentencesResources
    Network and pathway analysis of the associated proteins were performed using STRING v11.5
    STRING
    suggested: (STRING, RRID:SCR_005223)
    The data analysis and graphs were generated using GraphPad Prism (Ver 8.4.2).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 25 and 23. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

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  2. Version published to 10.1101/2021.10.21.465254 on bioRxiv